EXIT EXAM S16 A

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University of California, Davis *

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103L

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Feb 20, 2024

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MIC 103L Exit Exam (Form A) Page 1 MIC 103L Lab Exam (07 March 2016) • Please put your Name and Student ID Number on your ScanTron. • Please fill-in Form A . • Please mark your Answers to Multiple Choice Questions on your ScanTron. • Please erase any Mistakes on your ScanTron very Carefully.   Please do not ask any Questions during the Exam. ___________________________________________________________________ 1. A Researcher obtains a Soil Sample from a Garden. She uses a “Pinch” of this Soil to inoculate ~5.0 ml of Sodium Benzoate that has been added to each of five Test Tubes. She incubates these five inoculated Tubes for a Week at 25°C. Following Incubation all five Test Tubes appear Turbid. Our Researcher streaks a Loopful of turbid Sodium Benzoate from one of these five Tubes onto each of five Sodium Benzoate Plates. She incubates these five Sodium Benzoate Plates at 30°C for 48 Hours. After Incubation, Colonies of Bacteria appear on all of these Plates. Which of the Following represents the expected Appearance of the Colonies on these Plates and the expected Morphology and Gram Stain Results of the Bacteria taken from these Colonies? A. • Small Clear-to-White Colonies • Pink Rods. B. • Matte Construction Paper Colonies • Pink Rods. C. • Small Fluorescent Green Colonies • Pink Rods D. • Small Fluorescent Green Colonies • Purple Rugby-Ball-Shaped Rods

MIC 103L Exit Exam (Form A) Page 2 2. Our Researcher puts a “Pinch” of this same Soil Sample into a Test Tube containing ~5.0 ml of Water. She places this inoculated Test Tube in an 80°C Water Bath for 20 Minutes. After 20 Minutes she removes this Test Tube, allows it to cool, and vortexes this Test Tube. She then uses a Sterile Loop to streak a Sample from this Tube onto several Nutrient Agar Plates. After Incubation at 30°C for 48 Hours, Colonies of Bacteria appear on all of these Plates. Which of the Following represents the desired Appearance of the Colonies on these Plates and the expected Colony Morphology and Gram Stain Results of the Bacteria taken from these Colonies? A. • Small Clear-to-White Colonies • Purple Rods. B. • Matte Construction Paper Colonies • Purple Rods. C. • Matte Construction Paper Colonies • Pink Rods with Green Endospores D. • Small Clear-to-White Colonies • Purple Rods 3. Our Researcher uses a Patient Rectal Swab Plate to streak several EMB Plates. She incubates these EMB Plates at 37°C for 24 Hours. Colonies of Bacteria appear on all of these Plates. The Production of large Amounts of Acid results in the Precipitation of the metachromatic Dye Methylene Blue, making the Colonies themselves appear dark Purple. Which of the following represents the Expected IMViC Results for this Bacterium? Indole MR VP Citrate A. Yellow Yellow Red Redwoods Ring Layer Green _________________________________________________________________________________ B. Red Red Yellow Redwoods Ring Green _________________________________________________________________________________ C. Yellow Yellow Red Malibu Ring Layer Blue _________________________________________________________________________________ D. Red Red Yellow Malibu Ring Blue

MIC 103L Exit Exam (Form A) Page 3 Semi-Log Graph Paper

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MIC 103L Exit Exam (Form A) Page 4 4. Your Lab Partner obtains the following Data for the Bacterial Growth Curve Experiment: Time (Minutes) OD 0 0.052 10 0.092 20 0.160 30 Not Done 40 0.520 50 0.920  Semi-Log Graph Paper is on Page 3. What is the Doubling Time for this Bacterium? A. 12 Minutes B. 18 Minutes C. 24 Minutes D. 30 Minutes 5. The First Fundamental Step in Aligning your Microscope is to set the Working Distance between the Specimen and the Objective Lens. How is this done? A. You focus on the “P” Slide using the Fine and Coarse Focus Knobs. This raises and lowers both the Stage and the Condenser. B. You focus on the “P” Slide using the Fine and Coarse Focus Knobs. This independently raises and lowers the Stage (without moving the Condenser). C. You slowly close the Condenser Diaphragm until you see a Polygon. You focus on this Polygon using the Fine and Coarse Focus Knobs, which raise and lower both the Stage and the Condenser. D. You slowly close the Condenser Diaphragm until you see a Polygon. You focus on this Polygon using the Condenser Focus Knob, which independently raises and lowers the Condenser (without moving the Stage).

MIC 103L Exit Exam (Form A) Page 5 6. The Second Fundamental Step in Aligning your Microscope is to set the Working Distance between the Specimen and the Condenser Lens. How is this done? A. You use the Field Diaphragm Control to slowly close the Field Diaphragm until you see a Polygon. You focus on this Polygon using the Fine and Coarse Focus Knobs, which raise and lower both the Stage and the Condenser. B. You use the Field Diaphragm Control to slowly close the Field Diaphragm until you see a Polygon. You focus on this Polygon using the Condenser Focus Knob, which independently raises and lowers the Condenser (without moving the Stage). C. You use the Condenser Diaphragm Control to slowly close the Condenser Diaphragm until you see a Polygon. You focus on this Polygon using the Fine and Coarse Focus Knobs, which raise and lower both the Stage and the Condenser. D. You use the Condenser Diaphragm Control to slowly close the Condenser Diaphragm until you see a Polygon. You focus on this Polygon using the Condenser Focus Knob, which independently raises and lowers the Condenser (without moving the Stage). 7. The Third Fundamental Step in Aligning your Microscope involves creating an appropriately sized and centered Solid Cylinder of Light. How is this done? A. You use the Field Diaphragm Control to set the appropriate Size and the Condenser Centering Screws to center the Cylinder. B. You use the Condenser Diaphragm Control to set the appropriate Size and the Condenser Centering Screws to center the Cylinder. C. You remove an Eyepiece and use the Field Diaphragm Control to close the Field Diaphragm such that 80% of the Field of View is filled with Light. D. You remove an Eyepiece and use the Condenser Diaphragm Control to close the Condenser Diaphragm such that 80% of the Field of View is filled with Light.

MIC 103L Exit Exam (Form A) Page 6 8. The final Step in aligning your Microscope involves removing the Right Eyepiece and closing-down a Diaphragm so that 80% of the Field of View is Bright (or stated differently, so the outer 20% of the Field of View is Dark). How do you do this? A. • You use the Condenser Diaphragm Control. • The Condenser Diaphragm Control is the flat Black Plastic Lever that opens and closes the Condenser Diaphragm. • The Condenser Diaphragm itself is located inside the Condenser, just above the Condenser Lens. • Closing-down the Condenser Diaphragm trims the Cone of Light the Condenser is “condensing” onto the Specimen. • This reduces Glare and increases Contrast. B. • You use the Condenser Diaphragm Control. • The Condenser Diaphragm Control is the flat Black Plastic Lever that opens and closes the Condenser Diaphragm. • The Condenser Diaphragm itself is located inside the Condenser, just underneath the Condenser Lens. • Closing-down the Condenser Diaphragm trims the Cylinder of Light the Condenser is focusing onto the Specimen. • This reduces Glare but also reduces Contrast. C. • You use the Field Diaphragm Control. • The Field Diaphragm Control is the ribbed Plastic Ring on the Illumination System. • The Field Diaphragm itself is located just above the Illumination System in the Base of the Microscope • Closing-down the Field Diaphragm trims the Cone of Light leaving the Illumination System. • This reduces Glare and increases Contrast. D. • You use the Field Diaphragm Control. • The Field Diaphragm Control is the ribbed Plastic Ring on the Illumination System. • The Field Diaphragm itself is located inside the Condenser, just underneath the Condenser Lens • Closing-down the Field Diaphragm trims the Cylinder of Light leaving the Illumination System. • This reduces Glare but also reduces Contrast.

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MIC 103L Exit Exam (Form A) Page 7 9. Your Lab Partner did the Gram Stains of your Enteric Isolate (which is Enterobacter aerogenes ) and your Bacillus sp. Soil Isolate. Your Lab Partner makes a Smear of your Enterobacter aerogenes , allows this Smear to Air Dry for 20 Minutes and then Heat-Fixes this Smear. Your Lab Partner adds Crystal Violet and then goes out into the Hallway to answer a Text Message. After an Absence of nearly 10 Minutes your Lab Partner completes this Gram Stain, following the Instructions in the Lab Manual. Your Lab Partner makes a Smear of your Bacillus sp. on a separate Slide, allows this Smear to Air Dry for 20 Minutes and then Heat-Fixes this Smear. Your Lab Partner does all the Gram Stain Steps following the Instructions in the Lab Manual up to the Safranin Step. Your Lab Partner adds Safranin and then goes out into the Hallway to answer another Text Message. After yet another Absence of nearly 10 Minutes your Lab Partner completes this Gram Stain, again following Instructions in the Lab Manual. Which of the Following represents the Results of your Lab Partner ʼ s Gram Stains of your Enterobacter aerogenes and your Bacillus sp.? A. • Your Lab Partner ʼ s Enterobacter aerogenes will be Pink. • Your Lab Partner ʼ s Bacillus sp. will be Pink. ___________________________________________ B. • Your Lab Partner ʼ s Enterobacter aerogenes will be Purple. • Your Lab Partner ʼ s Bacillus sp. will be Pink. ___________________________________________ C. • Your Lab Partner ʼ s Enterobacter aerogenes will be Pink. • Your Lab Partner ʼ s Bacillus sp. will be Purple. _____________________________________________ D. • Your Lab Partner ʼ s Enterobacter aerogenes will be Purple. • Your Lab Partner ʼ s Bacillus sp. will be Purple.

MIC 103L Exit Exam (Form A) Page 8 10. A distant Uncle in Shanghai knows you are taking a Microbiology Lab and sends you a really spiffy Shenma Binocular “Biological Microscope” with no fewer than four (4) Objective Lenses on the Revolving Lens Turret. Golly! To your Surprise, your “Biological Microscope” has an Eyepiece Micrometer. It is marked from 0 through 100 Eyepiece Micormeter Units. You snaffle one of our MIC 103L Stage Micrometers to calibrate your Eyepiece Micrometer. When using the Yellow-Ringed 10X Lens you find that 25 Eyepiece Micrometer Units span 100 μ m on one of our Stage Micrometers. When using the White-Ringed 100X Oil Immersion Lens of your “Biological Microscope” a Vibrio cholerae measures Twenty (20) Eyepiece Micrometer Units. Approximately how large is this Bacterium? A. About 2.0 μ m. B. About 5.0 μ m. C. About 8.0 μ m. D. About 10.0 μ m. 11. You ʼ ve got your Bacteriophage in your “1” Dilution Tube. You notice that the Top Agar Tubes look suspiciously Full. ¡Canasta! ¡Crayola! ¡Sacramento! Your Lab Partner has measured 6.0 ml of Top Agar to each of the Top Agar Tubes in your 50°C Heating Block. What should you do? A. Continue the Experiment. This Set-Up will work and will give Data indistinguishable from the other Clans ʼ Data. B. Continue the Experiment. This Set-Up will work if you simply remember to divide the Number of Plaques you count by 2 (Two). C. Continue the Experiment. This Set-Up will work if you simply remember to multiply the Number of Plaques you count by 2 (Two). D. Stop the Experiment. This Set-Up will not work. Offer to commit Seppuku for the Honor of your Clan.

MIC 103L Exit Exam (Form A) Page 9 12. The other Clan at your Bench has just finished setting-up and labeling all the Tubes necessary for performing the Bacteriophage Titer Experiment. Uh-Oh! Someone in their Clan has added exactly 9.0 ml Tryptone Broth to each of their Dilution Tubes. Everyone in Lab is pointing at them and laughing. You step out into the Hallway, change into your Samurai O-Yori Battle Armor ( ), step back into the Lab and -- standing Arms akimbo -- announce -- “I ʼ m Samurai Microbiologist…” <<< Samurai Gong Sound Effect >>> “…And I am here to help you.” You ʼ ve definitely got this Clan ʼ s complete and undivided Attention now. What Advice should you give this Clan? A. This Set-Up will work as is and will give Data indistinguishable from the other Clans ʼ Data. B. • This Set-Up will work and will give Data indistinguishable from the other Clans ʼ Data if they simply transfer 200 μ l from each Dilution to each of the Top Agar Tubes. • No other Changes are Necessary. C. This Set-Up will work and will give Data indistinguishable from the other Clans ʼ Data if they: - Transfer 1000 μ l of Bacteriophage to their “1” (10 -1 ) Dilution Tube - Transfer 1000 μ l from each Dilution Tube to the next Dilution Tube in their Dilution Series (i.e. Transfer 1000 μ l from their “1” Tube to their “2” Tube…) - Transfer 100 μ l from each Dilution to each of the Top Agar Tubes. D. This Set-Up will work and will give Data indistinguishable from the other Clans ʼ Data if they: - Transfer 1000 μ l of Bacteriophage to their “1” (10 -1 ) Dilution Tube - Transfer 1000 μ l from each Dilution Tube to the next Dilution Tube in their Dilution Series (i.e. Transfer 1000 μ l from their “1” Tube to their “2” Tube…) - Transfer 200 μ l from each Dilution to each of the Top Agar Tubes.

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MIC 103L Exit Exam (Form A) Page 10 13. Your distant Uncle in Shanghai also sent you a Shenma Hemocytometer Your Shenma Hemocytometer has two Counting Chambers, each of which is divided into 9 Large Squares (1.0 mm x 1.0 mm; exactly like the 9 Large Squares in our Hemocytometers). The Large Square in the Middle of this Hemocytometer is subdivided into 25 Medium Squares (0.20 mm x 0.20 mm; 5 Squares across and 5 Squares down). Each Medium Square in turn is subdivided into 16 Small Squares (which you ignore). The Volume of the Counting Chamber of your Shenma Hemocytometer is identical to the Volume of the Counting Chamber of our Hemocytometers. You scoop-up of a small, isolated Yeast Colony to make a Yeast Suspension in exactly 5.0 ml of Sterile Saline and use this Suspension to load your Shenma Hemocytometer. The Center-most Square -- which fills your Field of View under 10X -- is further marked-off into 25 Medium Squares (0.2 mm x 0.2 mm). You count the Number of Yeast Cells in 5 of the 25 Medium Squares (the ones in each Corner, plus the one in the Center). Your Counts are: 42, 40, 36, 44 and 38. Approximately how many Yeast Cells were there in each Milliliter of this Yeast Suspension that you made? A. 6.4 x 10 6 Yeast Cells per Milliliter of Sterile Saline _____________________________________________________________ B. 1.0 x 10 7 Yeast Cells per Milliliter of Sterile Saline _____________________________________________________________ C. 3.2 x 10 7 Yeast Cells per Milliliter of Sterile Saline _____________________________________________________________ D. 5.0 x 10 7 Yeast Cells per Milliliter of Sterile Saline

MIC 103L Exit Exam (Form A) Page 11 14. You leave the Lab to get a Drink of Water and when you return, you are astonished to discover that your Lab Partner has both properly set-up the Dilution Tubes for the Viable Count Experiment and properly set-up the Dilution Series using the Hamburger Sample. Which of the following represents your Lab Partner ʼ s completed Dilution Series? A. All Six Dilution Tubes contain 4.5 ml of different Hamburger Dilutions. B. Five Dilution Tubes contain 4.5 ml of different Hamburger Dilution and one Dilution Tube contains 5.0 ml of a different Hamburger Dilution. C. Five Dilution Tubes contain 5.0 ml of different Hamburger Dilutions and one Dilution Tube contains 4.5 ml of a different Hamburger Dilution. D. All Six dilution Tubes contain 5.0 ml of different Hamburger Dilutions. 15. When you finish Viable Count Spread Plates, you tape your six MacConkey Agar Plates together in a Stack and incubate them for 24 Hours at 37°C, and you tape your six Nutrient Agar Plates together in a Stack and incubate them for 48 Hours at 30°C. Your Colony Counts for each Plate are as follows: MacConkey Agar Plates: 10 -3 10 -4 10 -5 10 -6 10 -7 10 -8 TNTC 513 51 5 0 0 Nutrient Agar Plates: 10 -3 10 -4 10 -5 10 -6 10 -7 10 -8 TNTC TNTC 213 22 2 0 Roughly what Percentage of the Colonies growing on your Nutrient Agar Plates were produced by Enteric Bacteria? A. None (0%) B. About One-Quarter (~25%) C. About Three-Quarters (~75%) D. All (100%)

MIC 103L Exit Exam (Form A) Page 12 MPN Table for Question 16 on Page 13 N o . Positive Tubes MPN 95% Confidence Limit 10 ml 1.0 ml 0.1 ml Lower Upper 0 0 1 3 < 0.5 9 0 1 0 3 < 0.5 13 1 0 0 4 < 0.5 20 1 0 1 7 1 21 1 1 0 7 1 23 1 1 1 11 3 36 1 2 0 11 3 36 2 0 0 9 1 36 2 0 1 14 3 37 2 1 0 15 3 44 2 1 1 20 7 89 2 2 0 21 4 47 2 2 1 30 10 150 3 0 0 23 4 120 3 0 1 39 7 130 3 0 2 64 15 380 3 1 0 43 7 210 3 1 1 75 14 230 3 1 2 120 30 380 3 2 0 93 15 380 3 2 1 150 30 440 3 2 2 210 35 470 3 3 0 240 36 1300 3 3 1 460 71 2400 3 3 2 1100 150 4800

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MIC 103L Exit Exam (Form A) Page 13 16. Early one Friday Morning you and your Lab Partner jump the Fence at Spunky Monkey ʼ s Jungle Gym & Watering Hole, swipe a 100 ml Water Sample from one of their disreputable Hot Tubs and then you each individually perform separate Most Probable Number (MPN) Tests. Your Lab Partner ʼ s Data from this MPN Test are as follows: Tube 1 Tube 2 Tube 3 10.0 ml (Red Lids) + + + _______________________________________ 1.0 ml (“M”) - - + _______________________________________ 0.1 ml (“C”) + + - You compare your Data with your Lab Partner ʼ s Data ( There’s an MPN Table on Page 12 ). After a few quick Manipulations of these Data you are able to determine that -- A. • There are approximately 1200 Coliform Bacteria per Liter of Spunky Monkey ʼ s Hot Tub Water. B. • There are just about exactly 12,000 Bacteria per Liter of Spunky Monkey ʼ s Hot Tub Water. C. • Your Lab Partner reversed the “M” and the “C” Labels. • Switch these Numbers and they match your Tubes. • Your Data indicate that there are approximately 150 Coliform Bacteria per Liter of Spunky Monkey ʼ s Hot Tub Water. D. • Your Lab Partner reversed the “M” and the “C” Labels. • Switch these Numbers and they match your Tubes. • Your Data indicate that there are just about exactly 15,000 Bacteria per Liter of Spunky Monkey ʼ s Hot Tub Water. 17. Which Biochemical Test is Positive if the Bacterium being tested has an Enzyme that breaks-down H 2 O 2 to O 2 + H 2 O? A. Oxidase B. Methyl-Red C. Catalase D. Citrate

MIC 103L Exit Exam (Form A) Page 14 18. Tetramethyl-p-Phenylenediamine turns Royal Thai Purple when -- A. Ammonia Salts have been broken-down to Alkaline Compounds that raise the pH. B. It is reduced by the Electron Transport Chain Enzyme Cytochrome B Reductase. C. It is oxidized by the Electron Transport Chain Enzyme Cytochrome C Oxidase. D. Ammonia Salts have been broken-down to Acidic Compounds that lower the pH. 19. Which Biochemical Test is Positive when the Bacterium being tested produces large Amounts of Stable Organic Acids? A. Indole B. Methyl Red C. Citrate D. Voges-Proskauer 20. Bromthymol Blue turns Deep Ocean Malibu Blue when -- A. Ammonia Salts have been broken-down to Alkaline Compounds that raise the pH. B. It is reduced by the Electron Transport Chain Enzyme Cytochrome B Reductase. C. It is oxidized by the Electron Transport Chain Enzyme Cytochrome C Oxidase. D. Ammonia Salts have been broken-down to Acidic Compounds that lower the pH. STOP! Fill-in “ Form A ” on your ScanTron. Double-check your Answers on your ScanTron. Turn-in your ScanTron. You may keep these Questions. The Archer’s Style during the Tournament will be remembered long after the Archer’s Score at the Tournament has been forgotten. - Confucius

EXIT EXAM S16 A

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